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免疫-PCR检测肾综合征出血热患者血清中抗核衣壳蛋白抗体方法的建立 被引量:3
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作者 史俊岩 牟玲 +6 位作者 罗恩杰 刘祥宁 王楠 张晓棠 吴云萍 赵新艳 林松 《中国人兽共患病杂志》 CSCD 北大核心 2004年第5期397-400,共4页
目的 建立高灵敏性和高特异性的免疫 -PCR方法检测肾综合征出血热 (HFRS)患者血清中抗核衣壳蛋白抗体。方法 用汉坦病毒 ( 76 - 118株 )重组核衣壳蛋白为抗原包被聚苯乙烯微滴度板 ,与经ELISA法和临床均已确诊为HFRS患者的血清进行抗... 目的 建立高灵敏性和高特异性的免疫 -PCR方法检测肾综合征出血热 (HFRS)患者血清中抗核衣壳蛋白抗体。方法 用汉坦病毒 ( 76 - 118株 )重组核衣壳蛋白为抗原包被聚苯乙烯微滴度板 ,与经ELISA法和临床均已确诊为HFRS患者的血清进行抗原抗体特异性反应 ,以链霉亲和素为桥梁将生物素标记的抗体与生物素标记的DNA指示分子相连 ,PCR扩增DNA指示分子 ,通过检测指示分子DNA来反映肾综合征出血热患者血清中抗核衣壳蛋白抗体的水平 ,并与传统的ELISA法进行比较。结果 免疫 -PCR检测肾综合征出血热患者血清中抗核衣壳蛋白抗体的敏感性是ELISA法的 2 5 0 0 0倍 ;30例HFRS阳性患者血清免疫 -PCR检测均为阳性 ,2 0例甲型肝炎阳性血清和 2 0例正常人血清免疫 -PCR检测均为阴性。结论 免疫 -PCR方法敏感性高 ,特异性强 。 展开更多
关键词 免疫-PCR 肾综合征出血热 抗核衣壳蛋白抗体
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Serial Expression of the Truncated Fragments of the Nucleocapsid Protein of CCHFV and Identification of the Epitope Region 被引量:8
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作者 Peng-fei WEI Yan-jun LUO +6 位作者 Tian-xian LI Hua-lin WANG Zhi-hong HU Fu-chun ZHANG Yu-jiang ZHANG Fei DENG Su-rong SUN 《Virologica Sinica》 SCIE CAS CSCD 2010年第1期45-51,共7页
The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen. Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV inf... The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen. Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections.In this study,expression vectors carrying series truncated fragments of the NP(nucleocapsid protein)gene from the S fragment of CCHFV strain YL04057 were constructed.The recombinant proteins were expressed in E.coli and purified for detection.The antigenic of the truncated fragments of NP was detected with a polyclonal serum(rabbit)and 2 monoclonal(mAbs)(14B7 and 43E5)against CCHFV by Western-blot analyses. The results showed that the three expressed constructs,which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum.The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein. 展开更多
关键词 Crimean-congo hemorrhagic fever virus (CCHFV) EXPRESSION EPITOPE Nucleocapsid protein (NP)
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